P-01
Clinical manifestation and diagnosis of invasive fungal sinusitis in 24 patients with hematological malignancy Masoud Mardani1, Yazdanali Faghani2, Mahdi Tabarraee3, Sara Abolghasemi1
1 Infectious Diseases and Tropical Medical Research Center, Shahid
Beheshti University of Medical Sciences,Tehran, Iran
2 Infectious Disease Specialist, School of Medicine, Islamic Azad
University of Tehran, Iran
3 Hematology and Oncology Center, Taleghani Hospital, Shahid Beheshti University of Medical Sciences, Tehran, Iran
Email: [email protected]
Introduction: Invasive fungal sinusitis (IFS) is a potentially lethal infection, especially in immunocompromised patients. The current study aimed to evaluate clinical manifestations, outcomes, and factors that may affect IFS in immonucompromised patients’ survival.
Materials and Methods: This cross-sectional, descriptive study was performed on patients admitted to Taleghani Hospital of Shahid Beheshti University of Medical Sciences, Tehran, Iran, during one year from October 2012. The clinical data of 24 patients with IFS were reviewed. All the patients had hematologic malignancies and had received broad-spectrum chemotherapy. Demographic data, clinical characteristics, presenting symptoms and signs, underlying diseases, and outcomes of the patients were studied.
Results: The age range of the patients was 15-60 years. IFS was identified as proven in 25%, probable in 66.7%, and possible in 8.3% of the cases. Serum galactomannan antigen was positive in 41.6% of the cases. In general, 15 out of 24 IFS patients had received antifungal chemoprophylaxis before diagnosis, while 54% of the patients received fluconazole and 8.3% itraconazole.
Aspergillus flavus (33%), A. fumigatus (20.8%), A. niger
(16.7%), and Mucor (16.7%) were responsible for incidence of IFS; moreover, 54% of IFS incidences occurred in summer. We found that 91.6% of IFS cases occurred during hospital construction, which was a risk factor in 91.6% of cases. Conclusion: Our study revealed that Aspergillus flavus was the most common fungal isolated pathogen in sinusitis. In addition,
Aspergillusfumigatus was the second isolated pathogen in the IFS patients. Hospital construction was an important environmental risk factor for acquisition of infection in hematological malignant patients. Additionally, the common causes of mortality in patients with IFS were primary disease and refractory to chemotherapy (37.5%).
Keywords: Sinusitis, Hematologic neoplasm, Mycoses,
Aspergillus, Mucor
P-02
Impact of stored grains contamination with Aspergillus species on farmer's lung disease
Seyed Reza Aghili1, 5, Ali Reza Khosravi2, Tahereh Shokohi3, Bahar Salmanian4
1 Department of Medical Parasitology and Mycology, Invasive Fungi Research Centre (IFRC), School of Medicine, Mazandaran University of Medical Sciences, Sari, Iran
2 Mycology Research Center, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran
3 Department of Medical Parasitology and Mycology, Invasive Fungi Research Centre (IFRC), School of Medicine, Mazandaran University of Medical Sciences, Sari, Iran
4 Department of Sciences, Seddighe-e Tahereh branch, Farhangian University, Sari, Iran
5 Department of Veterinary Medicine, Science and Research Branch, Islamic Azad University, Tehran, Iran,
Email: [email protected]
Introduction: Farmer's lung disease (FLD), also known as extrinsic allergic alveolitis, is an immunologically mediated inflammatory disease of the lung involving the terminal airways. Repeated inhalational exposure to biologic dusts including various Aspergillus species is one of the causes of this disease. Exposure to large quantities of contaminated wet agricultural products such as hay is the most common source of disease and
grain farmers usually are not at risk for the development of the disease. However, many types of mold, particularly Aspergillus
species, grow on insufficiently dried stored crops in granary. While working with moldy grains, the farmers inhale mold spores released as dust; inhalation of a large amount of this dust can be health risk for FLD.
Materials and Methods: For the purpose of data collection, 100 samples of paddy rice and 100 samples of polished rice collected from 100 granaries in Mazandaran, Iran, were studied for fungal contamination. For each sample, 24 grains were plated (12 per plate) on potato dextrose agar medium containing chloramphenicol (50 mg/l). All plates were incubated at 27°C for 7-10 days. Following incubation, the developing fungal colonies were counted and each morphologically unique fungal colony was sub-cultured and purified using standard techniques. The cultures were identified at genus or species level on the basis of macroscopic and microscopic features with the help of literature. With the aim of determining levels of fungal contamination of farmer’s stored rice in granaries, we discuss the potential risk of FLD in farmers and their households.
Results: The mortality rate of farmer's lung is reportedly 0-20%. Acute farmer's lung manifests as fever, chills, nonproductive cough, chest tightness, dyspnea, headache, and malaise. If the inhalational exposure is high, the patients may develop acute respiratory failure. The Aspergillus antigens, by humoral and cell-mediated immune responses, are responsible for the hypersensitivity reaction in FLD. Aspergillus colonization and infection must also be considered as potential triggers of airway symptoms in symptomatic farmers. Following inhalation of moldy grain, levels of interleukin-1 and interleukin-8 and tumor necrosis factor-alpha, as inflammatory mediators, were increased. In this study, the frequency rates of sample contamination with
Aspergillus species were 21.7% and 43.9% in stored paddy rice and polished rice, respectively. A. flavus (42.0%), A. niger
(32.4%), and A. fumigatus (11.9%) were the most frequent
Aspergillus species identified in 64/100 samples of paddy rice. However, A. flavus (57.9%), A. fumigates (17.5%), A. niger
(9.0%), and A. flavipes (7.1%) were the most isolated species in 61/100 samples of polished rice. Other species of Aspergillus
isolated from stored grain included A. oryzae, A. glaucus, A. nidulans, A. versicolor, and A. candidus.
Conclusion: Multiple factors are responsible for FLD. The evidence showed that in addition to genetic causes, inhalation and repeated exposure to Aspergillus species, particularly A. fumigatus, is one of the important causes of FLD in work environments as it activates adaptive response and leads to subsequent inflammation-driven lung damage. Prevention of fungal contamination of grain in the pre-harvest and post-harvest stages reduces humidity of granary. Therefore, using masks by farmers may reduce exposure with the amount of antigen. Keywords: Aspergillus, Farmer's lung disease, Fungal contamination, Inhalation, Antigen, Inflammation
P-03
Onychomycosis caused by Aspergillus species
Seyed Jamal Hashemi1, Mohsen Gerami shoar1, Roshanak Daie ghazvini1, Ensieh Zibafar1, Leila Hosseinpour1, Zeinab Borjian1, Kazem Ahmadikia1, Zahra Zareh1, Omid Raeesi1, Fereshteh Zarei1
1 Department of Medical Parasitology and Mycology, School of Public Health, National Institute of Health Research, Tehran University of Medical Sciences, Tehran, Iran
Email: [email protected]
of toenail infections. It is important to identify the causative agent of onychomycosis to ensure that the appropriate treatment is employed for each case. Therefore, the aim of this study was to determine the role and pattern of
Aspergillus species as the most common causative agent of onychomycosis due to NDM in Iran.
Materials and Methods: With the aim of determining the frequency of onychomycosis caused by Aspergillus species, a retrospective study was carried out during December 2014-September 2016 at the Medical Mycology Laboratory of Tehran University of Medical Sciences in Tehran, Iran. The results of direct and culture examination of 722 patients suspected of onychomycosis are presented in this study.
Results: According to the results, a total of 259 cases were diagnosed as onychomycosis out of which 85 cases (32.8%) were caused by NDM. Aspergillus species were the most frequently isolated etiological agent of onychomycosis due to NDMs (40.0%, N=34). Distributions of Aspergillus species were as follows: A. flavus (55.8%, N=19), A. niger (N=9), A. fumigatus
(N=2), A. terreus (N=1),Aspergillus species (N=1), A. candidus
(N= 1),and A.versicolor (N=1). Furthermore, the frequency of
Aspergillus species in females was 74.5% (26 cases). The mean patients age was 49 years. The most affected age group ranged between 50 and 73 years. The prevalence of toenail and fingernails onychomycosis were 84% and 16%, respectively. Distal and lateral subungual onychomycosis (DLSO) was the most clinical form of the infection. Among predisposing factors of onychomycosis, trauma, diabetes mellitus, and vascular disease were highlighted.
Conclusion: This study indicated that the frequency of onychomycosis due to Aspergillus species was 13.1%. Moreover,
A. flavus was found to be the most prevalent species. Consequently, the identification of the etiological agents of onychomycosis is necessary for appropriate therapy.
Keywords: Onychomycosis, non-dermatophytes molds (NDM), Aspergillus species
P-04
Disseminated Aspergillosis as the Herald Manifestation of Chronic Granulomatous Disease in an Adult Patient Ilad Alavi Darazam1, Davood Mansouri1, Hossein Akhavan Zanjani1, Davood Sanaee1
1 Institute of Tuberculosis and Lung Diseases, Shahid Beheshti University of Medical Sciences, Tehran, Iran
Email: [email protected]
Introduction: Chronic granulomatous disease (CGD) is an inherited defect in intracellular killing of ingested microorganisms, which are characterized as recurrent life-threatening bacterial and fungal infections, such as invasive aspergillosis, in early childhood.
Case Presentation: In this report, a 20-year-old girl, who was admitted due to low back pain, subcutaneous masses in the left thigh and left temporal part of scalp, as well as perihilar mass in the right lung, hypodense splenic lesions and heterogeneous infiltrative soft-tissue density in pelvic fosse, was assessed. Subcutaneous masses, bone marrow aspiration and spleen aspiration revealed septate, acute-angle hyphae in Gomori methenamine using silver staining and culture. The serum galactomannan was strongly positive, and a diagnosis of CGD was confirmed through nitroblue tetrazolium test (NBT= 0), demonstrating no dihydrorhodamine oxidation and abnormality. Results of this study were indicative of the delayed presentation of CGD until twenty years of age without previous infections and secondly, the high burden of Aspergillus infection involving multiple organs, particularly the isolation of this germ from the spleen and bone marrow. While Aspergillus infection in phagocytic disorders, especially CGD, is often presented as a as local slowly progressive infection, evaluation of our case revealed that multiple organs (skin, lung, pelvic cavity, spleen,
vertebrae and bone marrow) were affected. Another intriguing finding in this case report was the dramatic response to voriconazole, echinocandin and gamma interferon.
Conclusion: According to the results of this study, CGD must be considered in all patients with recurrent catalase-positive microorganism and fungal infections regardless of age, despite the fact that this condition usually manifests in children. Keywords: Disseminated Aspergillosis, CGD
P-05
Analysis of Aspergillus spp. spore load in indoor air samples of hospitals in Ardabil, Iran
Saeideh Amani Ghayum1, Sadegh Hazrati2, Farnaz Valedeiny Asl3, Solmaz Basiri Khiavi4
1 Department of Medical Mycology and Parasitology, Ardabil University of Medical Sciences, Ardabil, Iran
2 Department of Occupational and Environmental Health Engineering, School of Public Health, Ardabil University of Medical Sciences, Ardabil, Iran
3 Department of Vice Chancellor in Treatment Affairs, Ardabil University of Medical Sciences, Ardabil, Iran
Email: [email protected]
Introduction: Prevalence of aspergillosis is on the rise in hospitalized immunocompromised patients, often presenting as an invasive pulmonary disease. With the continuous increase in the number of severely immunocompromised patients, hospitals are faced with the growing rates of invasive aspergillosis and other opportunistic fungal infections. Considering their challenging treatment and fatal outcomes, preventive measures are of paramount importance in the control of invasive filamentous fungal infections. Until recently, inhalation of airborne Aspergillus conidia was believed to be the primary route of acquiring aspergillosis. Although efforts for the filtration of hospital air has led to the reduction of airborne conidia and frequency of invasive fungal infections, the association between the concentration of Aspergillus conidia in hospital air and risk of invasive fungal infections remains unclear.
Materials and Methods: Concentrations of airborne fungi were monitored during six months in different special care units of four hospitals in Ardabil, Iran. All airborne fungal samples were isolated from the air samples of four different hospital wards using a V-4 vacuum pump. To increase isolation accuracy, sample collection was performed using an active method. In addition, Czapek-Dox Agar containing chloramphenicol (40 mgl -1) incubated at the temperature of 30°C was used as the culture
medium. For sample discrimination, duplicate-slide culture assay was performed on PDA and CMA media. Moreover, microscopic examination was carried out using lactophenol cotton blue stain. Colony counts, CFU and other indices were analyzed statistically. Results: Out of 200 samples collected from different patient care environments, an average of 18 CFU/m3 was recovered at the temperature of 28°C and 35% humidity. In total, 57 species were identified as potential opportunistic fungi, including Aspergillus niger (55%), A. flavus (26.6%), A.fumigatus (8.3%), A. terreus
immunocompromised patients using a personal air-sampling device in intensive care units. However, no fungal sampler is currently available for this purpose.
Keywords: Aspergillus, Aspergillosis, Airborne conidia, Immunocompromised patients
P-06
Antifungal susceptibility profile of clinical and environmental Aspergillus flavus strains from Iran
Saham Ansari1, Mojtaba Taghizadeh-Armaki2,3,Mohammad T. Hedayati2,4, Paul E. Verweij,Seyedmojtaba Seyedmousavi5,6,7 1Department of Parasitology and Mycology, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran
2Department of Medical Mycology and Parasitology School of Medicine,
Mazandaran University of Medical Sciences, Sari, Iran
3Department of Medical Mycology and Parasitology, School of Medicine, Babol University of Medical Sciences, Babol, Iran
4Invasive Fungi Research Center, Mazandaran University of Medical Sciences, Sari, Iran
5Department of Medical Microbiology, Radboudumc, Nijmegen, The Netherlands
6Department of Medical Microbiology and Infectious Diseases, ErasmusMC, Rotterdam, The Netherlands
7Current Address: Laboratory of Clinical Infectious Diseases, National
Institute of Allergy and Infectious Diseases (NIAID), National Institutes of Health (NIH), Bethesda, Maryland, United States of America.
Email: [email protected]
Introduction: Aspergillus flavus has been frequently reported the leading cause of invasive aspergillosis in certain tropical and sub-tropical countries. The molecular epidemiology, and antifungal susceptibility profile of clinical and environmental A. flavus strain in Iran is poorly documented.
Material and methods: Two hundred A. flavus strains originating from clinical and environmental sources were phylogenetically identified at the species level using sequences of beta-tubulin gene. In vitro antifungal susceptibility testing was performed against seven antifungals by using the Clinical and Laboratory Standards Institute (CLSI-M38-A2) broth-microdilution method. Final concentrations of the following antifungal agents ranged from 0.016 to 16 mg/L: Amphotericin B (AMB), Isavuconazole (ISA), Itraconazole (ITC), Voriconazole (VRC), Posaconazole (POS), Caspofungin (CAS) and Anidulafungin (AFG).
Results: The geometric means of the MICs of the antifungals for all isolates were as reported below in increasing order; AFG 0.05, POS 0.11, ITC 0.20, CAS 0.35, VRC 0.57, ISA 0.98 and AMB 3.02 mg/L. The MIC ranges of antifungal agents were the following: AFG (0.016-0.125 mg/L), POS (0.016-4 mg/L), ITC (0.032-4.0 mg/L), CAS (0.016-0.5 mg/L), VRC (0.062-1.0 mg/L), ISA (0.032-1.0 mg/L) and AMB (0.1-4.0 mg/L), respectively.
Conclusions: Anidulafungin and posaconazole showed the greatest in vitro activity among systemic azoles and echinocandins, respectively. Antifungal susceptibility of A. flavus
was not linked with the clinical or environmental source of isolation.
Keywords: Antifungal susceptibility, Aspergillus flavus, Iran.
P-07
Invasive Aspergillus sinusitis successfully treated with voriconazole
Narges Aslani1, Ghasem Janbabaie2, Mahasti Babaeian2, Saman Rezaei3, Melika La’al Karghar4, Sahar Ghazanfari4, Nina Zahedi1, Hamid Badali5
1Student Research Committee, Mazandaran University of Medical Sciences, Sari, Iran
2Hematology & Oncology Research Center, Mazandaran University of Medical Sciences, Sari, Iran
3 Department of Otorhinolaryngology (ENT), Mazandaran University of Medical Sciences, Sari, Iran
4 Department of Mycology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran
5Invasive Fungi Research Center, School of Medicine, Mazandaran University of Medical Sciences; Sari, Iran
Email: [email protected]
Invasive Aspergillus sinusitis is a rare disease largely attributable to Aspergillus and Mucor species, widely recognized as a cause of morbidity and mortality in immunocompromised patients, with a high risk of neutropenia. Earliest indications of this disease in immunocompromised patients include fever, headache, nasal obstruction and facial swelling. However, this disease has a poor prognosis. This study aimed to describe a case of invasive
Aspergillus sinusitis successfully treated with voriconazole. A 22-year-old male patient presented with a history of acute lymphoblastic leukemia for nine months. A broviac catheter was inserted into the right subclavian vein, and the patient underwent chemotherapy with daunorubicin, cyclophosphamide and vincristine. After seven chemotherapy sessions, neutropenia (600/mm3) occurred in the patient. On day three of chemotherapy (session seven), while neutropenic, the patient developed a fever of 38.7°C with eye, throat and chills. All blood cultures were negative, and he had no history of tuberculosis, diabetes mellitus, asthma, corticosteroid use and long-term antibiotic therapy. Fiberoptic endoscopic sinus surgery was performed, and direct microscopic examination (10% KOH) revealed abundant septate fungal hyphae (dichotomous hyphae). A biopsy specimen was inoculated on Sabouraud dextrose agar (supplemented with chloramphenicol and brain-heart infusion agar) and incubated at temperatures of 25oC and 37oC for 48 h. Growth of yellowish colonies was observed, and the fungi were morphologically classified as Aspergillus flavus. Patient was started on intravenous ciprofloxacin, clindamycin, vancomycin, and liposomal amphotericin B (3 mg/kg/day for 5 weeks). Seven days after amphotericin B consumption, the patient developed fever (39.3°C) and chills. MICs of itraconazole, voriconazole and posaconazole were adjusted in accordance with the Clinical and Laboratory Standards Institute (M38-A2 document). Itraconazole had a low MIC value (0.25 µg/ml), followed by posaconazole (0.5 µg/ml), and voriconazole (0.5 µg/ml). Patient had a good response to voriconazole treatment (200 mg/day). However, despite antifungal therapy, patient conditions continued to deteriorate, and he died due to leukemia.
Keywords: Leukemia, A. flavus, Voriconazole
P-08
Highly Itraconazole Susceptible Aspergillus fumigatus and resistant Fusarium species Isolated from Avian Farms in Iran Narges Aslani1, Hamid Badali2
1Student Research Committee, Mazandaran University of Medical Sciences, Sari, Iran
2Invasive Fungi Research Center (IFRC)/Department of Medical Mycology and Parasitology, School of Medicine, Mazandaran University of Medical Sciences, Sari, Iran
Email: [email protected]
Methods: Environmental samples (nest material and compost heaps) from five nesting sites were collected and examined in terms of the growth of triazole-resistant A. fumigatus isolates. Cultures were prepared on a sabouraud dextrose agar plate, supplemented with 4 and 1mg/L of itraconazole and voriconazole, respectively, at 45°C for 72 h in the dark. All colonies growing on the plates, mimicking A. fumigatus
complexes, were sub-cultured. Strain identities were reconfirmed by DNA sequencing of the partial b-tubulin gene. Afterwards, in vitro antifungal susceptibility tests against triazole agents were performed based on the Clinical and Laboratory Standards Institute (CLSI) M38- A2 document.
Results: The MIC of itraconazole against A. fumigatus (15 strains) was less than breakpoint and epidemiological cut-off value, but Fusarium species (19 strains) had high MIC value (>16 µg/ml) which were resistant.
Conclusion: Although the number of azole-resistant isolates was limited, strict supervision of persistent environment azole-resistant isolates screening of azole resistance are vital to the development of approaches for the management of azole resistance in avian and human pathogenic fungi.
Keywords: Fusarium species, Aspergillus fumigatus, Avian Farms, Itraconazole
P-09
Streptomyces rochei as a sources of novel antifungal active compounds against azole-resistant Aspergillus fumigatus Sanaz Hadizadeh1, Hamid Forootanfar2, Gholam Hosain ShahidiBonjar3, Hamid Badali 4, Karamy Robati A1, Sahar amir rostami5, Seyyed Amin AyatollahiMousavi1
1Dept. of Medical Mycology & Parasitology, Faculty of Medicine, Kerman
University of Medical Sciences, Kerman
2Department of Pharmaceutical Biotechnology, Faculty of Pharmacy, Kerman University of Medical Sciences, Kerman
3Dept. of Plant Pathology & Biotechnology, College of Agriculture, Bahonar University of Kerman
4Department of Medical Mycology and Parasitology/Invasive Fungi Research Center (IFRC), Mazandaran University of Medical Sciences, Sari, Iran.
5Pharmaceutics Research Center, Institute of Neuropharmacology,
Kerman University of Medical Sciences, Kerman, Iran. Email: [email protected]
Introduction: Azole resistance in Aspergillus fumigatus is a major challenge and is emerging into a global health problem. Azole resistance may develop due to long terms of azole therapy or exposure of the fungus to the azole fungicides. Therefore, we aimed to evaluate the in vitro activity of Streptomyces rochei as a sources of novel antifungal active compounds against azole-resistant Aspergillus fumigatus
Material and Methods: In the present study, we report the isolation and characterization of actinomycetes compounds with a focus on the antifungal activity (minimum inhibitory concentration) against azole resistant A. fumigatus.
Results: Streptomyces rochei were isolated and confirmed by DNA sequencing (KP137826.1) and demonstrated antifungal activities against azole resistant A. fumigates with high efficacy. Conclusion: The observations from this study showed that these strains represent a particular interest for enlarged investigation, including isolation and characterization of pure active compounds and study of their biosynthesis.
Keywords: Streptomyces rochei, azole resistant, Aspergillus fumigates.
P-10
Aspergillus species in indoor environment of a burn hospital in Sari, Iran
Zeinab Bandalizadeh1, Mahin Tavakoli1, Afsane Vaezi1, Mohamad T. Hedayati1, Tahereh Shokohi1, Hamid Badali1 1Invasive Fungi Research Center, Department of Medical Mycology and Parasitology, School of Medicine, Mazandaran University of Medical Sciences, Sari, Iran
Email: [email protected]
Introduction: Nosocomial infections are mainly associated with high morbidity and mortality rates in healthcare systems. The presence of airborne fungi in hospital environments is of great concern due to their potential role as a source of hospital-acquired infections (HAI).Therefore, this study aimed to determine and compare the concentration of Aspergillus species in indoor environment of a burn hospital in Sari, Iran.
Materials and Methods: A cross-sectional analysis of
Aspergillus was performed in a burn hospital in Sari, using the quick take 30 pump-air sampler and carpet sterile fragments for sampling of air and surfaces in different wards of the hospital (e.g., operating room, intensive care unit, as well as surgery and burn wards, respectively). Collected samples were cultured on petri dishes containing Sabouraud dextrose agar with chloramphenicol. Plates were incubated at 27-30˚C for seven days. Grown Aspergillus spp. were identified in the level of species using morphological methods including macroscopic and microscopic characteristics.
Results: In this study, a total of 42 samples were collected and cultured. Mean level of recovered fungi was determined at 25.7 CFU/m3. Assessment of samples obtained from the air, surfaces and healthcare setting revealed that Aspergillus spp. were the most common fungi. Moreover, a total of 38 Aspergillus were isolated. Among the different Aspergillus species, A. fumigatus
(20.5%) was the most common, followed by A. flavus (17.9%),
A. niger (7.7%) and A. clavatus (1.3%), respectively. Conclusion: According to the results of this study, A. fumigatus
was spread as an important agent of nosocomial infections in hospital environments. It is recommended that contributing factors for the airborne fungal level in hospital environments be properly managed to minimize the risk of HAIs.
Keywords:indoor environment, burn hospital, Aspergillus spp.
P-11
Introduction of an Aspergillus polymerase chain reaction assay to clinical mycology service in Iran
Kambiz Diba1, Shahram Khademvatan1, Froozan Zarza2 1 Cellular and Molecular Research Center, School of Medicine, Urmia University of Medical Sciences, Urmia, Iran
2 Imam Educational Hospital, Urmia University of Medical Sciences, Urmia, Iran
Email: [email protected]
Introduction: Aspergillus species are abundant and widely distributed in soil, water, air, seed, and food. These species are associated with allergic bronchopulmonary disease, mycotic keratitis, otomycosis, nasal sinusitis and invasive infection. In this study, we developed a polymerase chain reaction-single-strand conformational polymorphism method (PCR-SSCP) to identify the most common Aspergillus species.
Materials & Methods: The current study utilized Aspergillus
clinical isolates of an educational hospital in Urmia, Iran, as well as some Aspergillus standard species obtained from Japan Collection of Microorganisms. All Aspergillus isolates were identified, using the morphological (colonies and microscopic) features. For the molecular identification, the ITS2 region of rDNA gene (approximate length size: 330 bp) was amplified in PCR. The PCR product was incubated at 95ºC for 5 min, and then moved quickly into ice bath for immediate quenching. A vertical electrophoresis with 6-12% gradient poly-acrylamide gel was used full time cooling at 4ºC.
Results: The tested Aspergillus species including A. nidulans, A. fischeri, A. fumigatus, and A. niger discriminated. Consequently, SSCP assay enabled us to identify above Aspergillus species within 8-12 h after overnight incubation.
recommend this method as a compliment test to be used with other molecular methods such as PCR-restriction fragment length polymorphism in order to identify more Aspergillus species. Keywords: Rapid identification, Aspergillus, Clinical case
P-12
Aspergillus identification in clinical and hospital sources, Urmia, Iran
Kambiz Diba1, Davood Jabari2, Elnaz Hosseianlipoor2
1 Cellular and Molecular Research Center, School of Medicine, Urmia University of Medical Sciences, Urmia, Iran
2 Department of Mycology and Parasitology, School of Medicine, Urmia University of Medical Sciences, Urmia, Iran
Email: [email protected]
Introduction: In spite of a low percent (1%) of fungal hospital acquired infections, Aspergillus species are the main agents of fulminate fungal infections. This study aimed to detect the sources of nosocomial infections through the identification of
Aspergillus species isolated from hospital, clinical and environmental samples.
Materials and Methods: Samples in this study included
Aspergillus isolates with confirmed hospital-acquired infections and environmental specimens collected from air, swabs of the walls, curtains, beds, blankets, trolleys, air conditioners and medical devices. A morphological diagnosis was performed on Sabouraud glucose agar and Czapek dox agar for the identification of fungal agents, which was confirmed by the PCR-RFLP method. In addition, random amplified polymorphic DNA (RAPD) technique was performed based on the polymerase chain reaction (PCR) for a molecular typing of environmental isolates. Results: In this study, the final results revealed the presence of
Candida spp., Aspergillus spp. and other fungi in 110 fungal isolates. Among the clinically isolated Aspergillus, the most frequent species were A. flavus (47%), A. fumigatus (29.4%) and
A. niger (23.6%), respectively. Environmental specimens contained Aspergillus isolates as follows: A. niger (43.7%), A. flavus (41.8%) and A. fumigatus (14.7%). Comparison of clinical and environmental isolates demonstrated RAPD settings with similar patterns for the two clusters.
Conclusion: According to the results of this study, while molecular typing of Aspergillus species by PCR-RFLP and RAPD might be a simple method to find hospital Aspergillus
sources, the final results are mostly inaccurate.
Keywords:Aspergillus species, hospital sources, RAPD-PCR, PCR-RFLP
P-13
Survey of fungal contamination in hospital wet cooling systems in Arak city, Iran
Mojtaba Didehdar, Mahdi Mosayebi, Zahra Eslamirad, Reza Hajihossein
Department of Medical Mycology and Parasitology, School of Medicine, Arak University of Medical Sciences, Arak, Iran
Email: [email protected]
Introduction: Fungal infections are common complication during hospitalization, especially in immunocompromised patients. Wet cooling systems in hospitals are considered as a major source of nosocomial infections. This study aimed to evaluate the wet cooling systems of hospitals in terms of fungal contamination in Arak city, Iran.
Materials and Methods: This descriptive, cross-sectional study was conducted during May-September 2016. Samples were randomly collected from the water and straws of 84 wet cooling systems in four hospitals in Arak, Iran. Samples were cultured in Sabouraud dextrose agar containing chloramphenicol. Identification of fungi was performed using the slide culture method.
Results: Out of 84 wet cooling systems, fungal contamination was detected in 32 cases (38.1%). The highest rate of fungal contamination was observed in oncology wards and coronary care
units. Moreover, the most frequent fungi isolates in the selected hospitals were Aspergillus spp. and Candida spp., respectively. Conclusion: According to the results of this study, significant fungal contamination was present in the hospital wards using wet cooling systems, particularly Aspergillus contamination. Therefore, it is recommended that non-aqueous or closed-cycle air conditioning systems be used in hospitals, especially in the care environment of susceptible patients.
Keywords: Fungi, Wet cooling system, Hospital, Arak
P-14
Use of CSP typing for genotyping azole-resistant and susceptible Aspergillus fumigatus isolates in Iran
Mahsa Falahati1, Hamid Badali1, Hamed Fakhim1, Saham Ansari1, Mahdi Abastabar1
1Invasive Fungi Research Center, Mazandaran University of Medical
Sciences, Sari, Iran
Email: [email protected]
Introduction: Aspergillus fumigatus is globally known as a leading cause of various clinical diseases, accounting for an alarmingly high mortality rate in immunocompromised hosts. Recognition of pathogen dispersion and relatedness is essential for determining the epidemiology of nosocomial infections and designing effective pathogen control techniques. This study aimed to investigate the diversity and putative origins of clinical and environmental susceptibility and resistance of A. fumigatus
isolates in Iran.
Materials & Methods: In total, 79 A. fumigatus isolates were evaluated, including 15 azole-resistant and 64 azole-susceptible isolates, which were genotyped using partially cell surface protein (CSP) gene.
Results: Seven distinct repeat types (r01, r02, r03, r04, r05, r06, and r07) and 11 different CSP variants (t01, t02, t03, t04A, t06A, t06B, t08, t10, t18A, t18B, and t22) were observed among the isolates. Interestingly, t06B, t18A and t18B were exclusively found in azole-resistant A. fumigatus isolates (TR34/L98H or non-TR34/L98H). Simpson’s diversity index (D) was calculated at 0.78.
According to the results, isolates with significant resistance were genetically less diverse compared to azole-susceptible isolates. However, azole-resistant A. fumigatus without TR34/L98H were more diverse than azole resistant with TR34/L98H. In addition, the relatively close genetic relationships and limited CSP type diversity of the TR34/L98H isolates, compared to those of the azole-susceptible wild-type isolates, indicates that the independent and repeated emergence of the TR34/L98H mechanism to be unlikely.
Conclusion: It seems that resistant A. fumigatus, which has been emerging across the world, could spread easily through producing a large number of asexual airborne conidia. Furthermore, it has been suggested that CSP types might have a common ancestor that developed locally, migrating to different regions of the world.
Keywords: CSP typing, Aspergillus fumigatus, Azole-resistant and susceptible, Iran
P-15
Evaluation of antifungal activity of graphene oxide conjugated with indolicidin against Aspergillus spp. and Candida albicans
Ali Farzanegan1, Maryam Rudbary1, Mehraban Falahati1 1 Department of Parasitology and Mycology, School of Medicine, Iran University of Medical Sciences, Tehran, Iran
Email: [email protected]
interest. This study was performed to determine antifungal activity of graphene oxide conjugated with indolicidin as compared to amphotericin B and fluconazole on standard strains of C. albicans and Aspergillus spp.
Materials and Methods: Graphene oxide (GO) was synthesized using Homer method and carboxyl groups were increased on it by C2H3BrO2 (Bromoacetic acid) and sodium hydroxide; thereafter, it was activated by dimethylaminopropyl chloride hydrochloride (EDC) and N-hydroxysuccinimide (NHS). Then the cationic antimicrobial peptide (indolicidin) was added to graphene for nanocomposite synthesis, which was confirmed by FTIR , NMR , TGA, and SEM analysis.For minimum inhibitory concentration (MIC) assessment, C. albicans (ATCC 10231) was cultured on Sabouraud dextrose agar (SDA) and incubated at 35°C for 48 hours. Moreover, A. fumigatus (ATCC204305), A. flavus (CBS 625166), and A. niger(ATCC 1105 ) were cultured on Czapek Dox Agar and incubated at 30°C for one week. Fungal suspensions were prepared at concentration of 1×103 (CFU/ml). MIC and minimum fungicide concentration (MFC) were determined by microdilution broth method with ranges 200-0.39 μg/m for nanocomposite, 128-0.25 μg/ml for fluconazole, 100-0.19 μg/ml for IN, 200-0.39 μg/ml for GO, and 32-0.06 μg/ml for amphotericin B. Each test was carried out in triplicate. In addition, negative and positive controls were considered. Results: Our results indicated that nanocomposite had strong inhibitory effect against C. albicans (MIC: 3.12 μg/ml) compared to indolicidin and GO alone. Furthermore, at concentration of 25 μg/ml it had candidacidal activity, whereas this nanocomposite did not show any inhibitory effect on Aspergillus spp. Conclusion: Designing a new drug delivery system via nanotechnology, which is able to inhibit fungal growth, can be one of the drug delivery system objectives for improving treatment with minimum side effects. According to our results, this nanocomposite with suitable MIC against C. albicans can introduce an appropriate component for inhibition of candida growth. The integrity of Aspergillus cell wall may explain the resistance of Aspergillus to nanocomposite penetration. However, further in-vitro and in-vivo studies should be performed on the safety and detailed mechanisms of this nanocomposite. Keywords: Graphene oxide, Indolicidin, MIC, Nanocomposite,
Candida, Aspergillus
P-16
Isolation of Aspergillus species from Educational hospitals in Ahvaz, Iran
Mahnaz Fatahinia1, Neda Kiasat1, Ali Zarei Mahmoudabadi1,2 1 Department of Medical Mycology, School of Medicine, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran
2 Health Research Institute, Infectious and Tropical Diseases Research Center, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran
Email: [email protected]
Introduction: Nosocomial aspergillosis is a common complication among immune-compromised individuals and high-risk patients. Aspergillosis has become a major invasive fungal infection in hospitals in recent decades. Aspergillus, a saprophytic fungus with wide distribution, is associated with systemic infections in predisposed patients with a mortality rate of about 85%. Aspergillus speices produce microscopic conidia, which can easily spread through the air. These conidia can enter alveolar through the airway and may cause different types of aspergillosis. This study aimed to evaluate diversity and abundance of
Aspergillus in air samples of educational hospitals in Ahvaz, Iran. Materials and Methods: The air was sampled from seven wards of five hospitals using Quick Take 30 Sample Pump (SKC, USA). As a result, a total of 175 air samples were collected, and then analyzed. Species of Aspergillus were identified, using macroscopic and microscopic features. Finally, the number of colonies forming units per cubic meter of air (CFU/m³) was calculated:
𝐶𝐹𝑈/𝑚3 = 𝑐𝑜𝑙𝑜𝑛𝑖𝑒𝑠 𝑛𝑢𝑚𝑏𝑒𝑟
𝐷𝑒𝑏𝑏𝑖𝑒(28.3) × 𝑇𝑖𝑚𝑒(min)
Results: According to the results of the study,420 colonies of
Aspergillus species were detected. The highest and least concentration of conidia in the air was related to surgery (1366 CFU/m3) and burn wards (412 CFU/m3), respectively. The most abundant Aspergillus species were as follows: A. niger (17.6%),
A. terreus (15%), A. fumigatus (11.4%), and A. flavus (8.3%). Conclusion: The World Health Organization (WHO) has suggested limit of 50 CFU/m3 for fungi in the hospital air. Regarding this, this study demonstrated that the contamination level is too high. Considering the presence of Aspergillus species in the indoor hospital as well as the resistance of A. terreus to amphotericin B in vivo and in vitro, monitoring is necessary to prevent possible hospital infections.
Keywords:Aspergillus, Hospital, Air, Nosocomial
P-17
Study of hemolysin gene "aspHS" and its phenotype in Aspergillus fumigatus isolates
Farzaneh Ganj1, Majid Zarrin1
1 Department of Medical Mycology, School of Medicine, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran
Email: [email protected]
Introduction: Aspergillus fumigatus is the most pathogenic of all aspergilli fungi, with more than 90 percent of mortality rates of diseases. The major cause of this virulence is pathogenic factors dependent on specific gene sequences. Such a factor is the release of haemolysin which is encoded by aspHS gene. Haemolysin helps fungi to kill blood cells and has cytotoxic effects on endothelial cells and macrophages. Diagnosis merely based on morphological properties provides difficulties and is prone to uncertainties. Molecular biological methods with higher speed and precision are however able to differentiate morphologically identical genus, and help with detection of genotypes and polymorphs. In vitro studies indicate that the path of haemolysin release is different in diverseA. fumigatus isolates. The present study provides a phenotypic examination of haemolysin enzyme in these isolates through real-time PCR-RRLP to detect differences in aspHS gene sequence.
Materials and Methods: Fifty three A. fumigatus isolates including 4 standard isolate, 10 clinical, and 39
environmental isolates were approved morphologically for study. Due to higher resolution in haemolytic activity in A. fumigatus, 10 A. niger isolates were selected as control group. To measure haemolytic activity, isolates were incubated in blood agar medium at 37 °C for 48h. F-Asphs and R-Asphs primers in PCR was able to obtatin a 180bp band and Afhem2 and Afhem1 primers recovery a 450bp band. Restriction enzymes NcoI and
TaqI also were able to identification of different genotypes. AnoR and AnoF primers, redesigned according to A. niger haemolysin sequences, provided was able to identification of wider sequences in A. fumigatus haemolysin.
Results: A phenotypic comparison of haemolysin activity in A. fumigatus and A.niger indicated that all isolates were able to create haemolytic cover with varying degrees (6 to 7.6) around blood agar colonies. PCR products from sequence were strikingly similar (99 percent) to haemolysin in data base. After digestion of PCR products by restriction enzymes NcoI and TaqI, a similarity of 97 percent also was shown, with only 3 percent polymorphism. Using haemolysin sequences in A. niger and its primers, and AnoF and AnoR primers, a band were detected for
Conclusion: The present study indicated that despite little polymorphism in haemolysin gene, it is still a proper sequence for genetic marking of A. fumigatus.
Key words: Aspergillus fumigatus, RCR-RFLP, aspHS
P-18
In vitro susceptibility testing of Aspergillus species against five antifungal agents
Parisa Badiee1, Fatemeh Ghasemi2, Hadis Jafarian1,Seyed Ahmad Ali Dastgheib1
1 Alborzi Clinical Microbiology Research Center, Shiraz University of Medical Sciences, Shiraz, Iran
2Shiraz Islamic Azad University, Shiraz, Iran
Email: [email protected]
Introduction: The incidence of serious infections caused by opportunistic fungi has increased dramatically in immunocompromise patients. Aspergillus species are the most common etiologic agent. The aim of this study was to determine the in vitro activities of five antifungal agents, namely amphotericin B, voriconazole, itraconazole, posaconazole, and caspofungin, against Aspergillus species isolated from patients. Materials & Methods: For the purpose of data collection, a total of 68 Aspergillus specieswere collected from the patients. All samples were plated on Sabouraud 4% dextrose agar and identified by routine and restriction fragment length polymorphism methods. Susceptibility tests were performed for five antifungal agents, using Clinical and Laboratory Standards Institute M38-A2 microdilution reference method. Data were analyzed by SPSS version 18.
Results: The collected species included A. flavus (33), A. fumigatus (29), A. niger (4), and A. species(2). The MIC 50 and MIC 90 values for amphotericin B, voriconazole, itraconazole, posaconazole, and caspofungin were found to be 2.0 and 8.0 μg/ml; 0.25 and 0.75 μg/ml; 0.03 and 1.5 μg/ml; 0.03 and 0.13 μg/m, and 10.03 and 0.50 μg/ml, respectively.
Conclusion: Regarding the high MIC 90 value for amphotericin B and itraconazole, the effective antifungal agents for treatment of Aspergillus infections in our region were voriconazole, posaconazole, and caspofungin.
Keywords: Aspergillus fumigatus, Aspergillus flavus, Voriconazole,Amphotericin B
P-19
Iatrogenic inter-abdominal and injection site abscess formation due to Aspergillus fumigatus in two patients: A case report
Nahid Gholineghad1, Tahereh Shokohi2, Seyed Reza Aghili2, HossseinKarami 3, hassan Asghari lamoki 4, Zahra Seifi1, Zeynab Bandalizadeh1, Somayeh Alizadeh5
1 Student Research Committee, School of Medicine, Mazandaran University of Medical Sciences, Sari, Iran
2Invasive Fungi Research Center (IFRC), Mazandaran University of Medical Sciences, Sari, Iran
3Thalassemia Research Center, Mazandaran University of Medical Sciences, Sari, Iran
4 Specialist in Infectious Diseases & Tropical Medicine, Zare Hospital,
Sari, Iran
5 Boali Sina Hospital, Sari, Iran
Email: [email protected]
Introduction: Aspergillosis is an opportunistic infection with a very high mortality rate, which especially occurs in diabetic, neutropenic, and immunosuppressive patients. The cutaneous aspergillosis is a rarely encountered form of aspergillosis that may occur as either primary or secondary infection. In primary cutaneous aspergillosis, the lesion occurs as a result of direct inoculation of Aspergillus spores at the site of injury following intravenous catheter, trauma, occlusive dressings and tapes, burns, or surgery. Among Aspergillus species, the most common causative agent of opportunistic infections in humans is A. fumigatus. Herein, we present two rare cases of iatrogenic
Aspergillus abscess formation following neglected left foreign body and indwelling angiocatheter.
Case Presentation
Case 1: A 4-year-old girl with a history of acute lymphoblastic leukemia was diagnosed with high fever and abscess in injection site of catheter into her right hand. The specimen was obtained from aspiration of the abscess and stained with Calcofluor White for direct microscopy. Dichotomous branching septate hyphae were observed in direct examination. Culture of the purulent exudates was performed on Sabouraud dextrose agar plates and fungal colonies were grown after five days at 30°C. Aspergillus fumigatus was identified using morphological characterization and confirmed by molecular method. Treatment with voriconazole was administered and the patient’s signs improved. Case 2: A 45-year-old woman with a history of type II diabetes was diagnosed with chronic wound with purulent exude localized on suture line after gynecological surgery. Radiographic imaging showed foreign object and exploratory laparotomy revealed unintentional retained surgical sponge in the patient's abdomen during a surgical procedure a week previously. With sampling from the abscess in site of suture and then staining with Calcofluor White, dichotomous branching septate hyphae were observed in direct examination. The purulent exudates were cultured on Sabouraud dextrose agar plates. After growth of fungal colonies, A. fumigatus was identified using morphological characterization and molecular method. The patient’s signs improved after treatment with voriconazole.
Conclusions: Cutaneous aspergillosis may manifest with non-specific cutaneous lesion and abscess presented by swelling and redness and exudes as an iatrogenic complication following a common care and surgical procedure. These cases are being presented to increase awareness of clinicians and pathologists regarding the fact that cutaneous aspergillosis could present as a chronic abscess that should be diagnosed accurately.
Keywords: Aspergillus fumigatus, abscess formation
P-20
Fungal rhinosinusitis in immunocompromised patients Parisa Badiee, Hadis Jafarian
Alborzi Clinical Microbiology Research Center, Shiraz University of Medical Sciences, Shiraz, Iran
Email: [email protected]
Introduction: Fungal rhinosinusitis can cause fatal infections in immunocompromised patients. In this study, we investigated the etiology of such infections among immunocompromised patients admitted to Nemazee Hospital, Shiraz, Iran.
Materials and Methods: In general, 67 patients with suspected rhinosinusitis, with underlying diseases such as diabetes, organ transplantation, and hematological disorders were entered into this study. Tissue samples from sinuses were cultured on Sabouraud dextrose agar. The isolated fungi were identified by routine mycology lab methods. Pathology and computed tomography scan results were collected from patient records. Results: A total of 36 patients had documented infections by histopathology smear, and fungi were isolated from 18 patients (18/39, 46.2%). The isolated agents were 10 Mucoraceae, 13
Aspergillus flavus, 1 Aspergillus fumigates, and 4 Candida spp. Conclusion: Aspergillus spp. is one of the most frequently isolated fungi from infected patients. Accurate and early diagnosis is very important for successful treatment, especially in immunocompromised patients.
P-21
Relationship between aflatoxin exposure and low birth weight
Aghili Seyed Reza 1, Javad Javidnia2, Bahar Salmanian3 1 Assistant Professor, Faculty Member of Department of Medical Parasitology and Mycology /Invasive Fungi Research Centre (IFRC), School of Medicine, Mazandaran University of Medical Sciences, Sari, Iran
2 PhD student of Medical Mycology, Student Research committee, School of Medicine, Mazandaran University of Medical Sciences, Sari, Iran 3 Faculty Member of Department of Science, Farhangian University, Department of Sciences, Sari, Iran
Email: [email protected]
Introduction: Low birth weight (LBW) is a term used to describe babies who are born weighing less than 2,500 grams. LBW is a risk for children's health and a problem of public health in under developing countries. Prevalence of LBW based on Iranian hospital data in 2013, was stimated7% and the evidence suggests that it is on the rise during two decades. Poor-quality diets and high rates of infection in pregnancy result LBW, but the relative contributions to this subject are unknown.This study tried to show information available on the potential impact of exposure to mycotoxins, including aflatoxin in the incidence of LBW. Materials & Methods: In this retrospective study, we did an extensive literature review of published studies about mycotoxins particularly aflatoxin and low birth weight and infant growth. Results: Aflatoxins are toxic metabolites produced by certain fungi (Aspergillus flavus and Aspergillus parasiticus) in/on foods and feeds. Aflatoxins are a cause of hepatotoxicity, cancer and in high doses, have caused deaths from aflatoxicosis. More recently, have been reported that there were significant negative effects of aflatoxin on child growth, as well as immune modulation in prenatal and after birth. These observations are consistent with impaired fetal development, immune deficiency and gut dysfunction in animal models. Many studies suggest that aflatoxin exposure contributes to stunting, independent of and with other risk factors (About 27% impact) and high exposure aflatoxin during pregnancy lead to fetal growth restriction and LBW. A study in The Gambia found a significant association between in utero aflatoxin exposure and growth faltering in infants.
Conclusion: Our study concludes that surveillance information on exposure to mycotoxins such as aflatoxins are generally lacking outside in the development countries such as Iran. Available data from measurements of contaminated crops and through the use of disposal biomarkers in exposed society indicate that mycotoxin exposures can be high throughout development countries, as well as in Iran and other parts of Asia. However, the validity of the findings from studies on aflatoxin and LBW is uncertain because they have small sample sizes for adverse birth outcomes, and thus may not be sufficiently powered to detect important outcomes. So, to confirm this issue, need further investigation.
Key words: Mycotoxin, Aspergillus, Aflatoxin, Food contamination, Low birth weight
P-22
Epidemiology and microbiology of fungal diseases: A survey in South of Iran
Azadeh Karami Robati, Seyyed Amin Ayatollahi Mousavi, Sanaz Hadizadeh
Department of Mycology and Parasitology, Afzalipoor Faculty of Medical Sciences, Shahid Bahonar University of Kerman, Kerman, Iran
E-mail: [email protected]
Introduction: Invasive fungal infections are a growing public health problem and cause high morbidity and mortality rates in the nosocomial setting.
Materials and Methods: In a two-year retrospective study of over 248 transplantation patients, the incidence of community-acquired fungal infections in three hospitals of Kerman, Iran was
investigated. Isolated molds were identified based on colonial morphology and slidecultures. Yeast colonies were detected by growth on Corn Meal Agar (CMA) and PCR- RFLP. Community-acquired infection isolates from 127 (70.5%) patients were obtained.
Results: Candida spp. were the most commonly isolated invasive yeasts, and Aspergillus spp. were the most commonly (15%) isolated invasive moulds. C. albicans was the most common (51.8%) microorganism, followed by C. glabrata, C. parapsilosis, C. kafier. A. fumigates remains the most frequently isolated mold, followed by A. niger.
Keywords: Community-acquired fungal infections, Aspergillus
spp., Candida spp.
P-23
Morphological identification of Aspergillus species in Zahedan
Nasser Keikha1, Ali Jalali 2, Mohadese Shahraki 2 , Nasimeh Marghzari1, Sanaz Aghaei gharebolagh3, Bahman Fouladi4 1 Infectious Diseases and Tropical Medicine Research Center, Zahedan University of Medical Sciences, Zahedan, Iran
2 Islamic Azad University of Zahedan, Zahedan, Iran
3 Department of Medical Mycology and Parasitology, School of Public
Health, Tehran University of Medical Sciences, Tehran, Iran
4 Department of Medical Mycology and Parasitology, Zabol University of
Medical Sciences, Zabol, Iran
Email: [email protected]
Introduction: Aspergillus species are saprophytic fungi. Since these species are ubiquitous in the environment, people may inhale hundreds of Aspergillus conidia per day.
Aspergillus conidia can cause a variety of clinical manifestations depending on the immune status of the host. The present study aimed to identify Aspergillus species isolated from environmental samples, using morphological (macroscopic and microscopic) characteristics.
Materials & Methods: This descriptive study employed two differential media, namely Czapek Dox agar (CZA) and Sabouraud glucose agar (SGA) with chloramphenicol for air sampling. Air samples were collected by placing the media plates in the exposure of air flow for 15 min at a height of 120 cm from the ground in five areas of Zahedan, Iran. After seven days of incubation, the plates (in triplicates) were observed for macroscopic characteristics such as colony diameter, exudates, and colony reverse and microscopic characteristics including conidiophores, vesicle, metulae, phialides, and conidia. For microscopic characteristics slides were stained with lectophenol cotton blue.
Results: According to the results of the study, a total of 330 fungal isolates were obtained. Themost common Aspergillus
species were demonstrated to be A. fumigatus (47.5%), A. niger
(40.5%), and A. flavus (18%), respectively.
Conclusion: Our findings demonstrated that morphological methods are useful to identify the species of Aspergillus; however, molecular techniques are improved and become highly available.
Keywords:Aspergillus, Morphological identification, Zahedan
P-24
Molecular characterization of Aspergillus niger and Aspergillus flavus isolated from Tehran air using RAPD-PCR Firoozeh Kermani1, Masoomeh Shams-Ghahfarokhi2, Seyed Reza Aghili1, Mehdi Razzaghi-Abyaneh3, Samira Dodangeh1 1 Department of Medical Mycology and Parasitology, Mazandaran
University of Medical Sciences, Sari, Iran
2 Department of Mycology, Faculty of Medical Sciences, Tarbiat Modares
University, Tehran, Iran
3 Department of Mycology, Pasteur Institute of Iran, Tehran, Iran Email: [email protected]
patients. Aspergillus spores exist in the indoor and outdoor air; therefore, they can easily reach the respiratory systems. The antifungal drug resistance is currently the subject of detailed investigations and reviews. Regarding this, the use of molecular characterization for identification of Aspergillus species in the inhaled air is very important and usually random amplification of polymorphic DNA polymerase chain reaction (RAPD-PCR) technique is used for detecting the genetic variability.
Materials and Methods: For the purpose of data collection, 18
Aspergillus isolates were collected including two species of A. niger and A. flavus. Out of the collected samples, 11 isolates were
A. niger obtained from the air of districts 1, 4, 8, 11, and 18 and seven isolates were A. flavus obtained from the air of districts 2, 3, 19, and 22 of Tehran Municipality. The two species were subjected to RAPD-PCR using 7 primers for determining the molecular characterization and fingerprint patterns.
Results: The dendrogram obtained from the data by 7 random decamer primers showed that 11 isolates of A. niger had 41 % similarity to each other and they were resided in one group with three branches. However, 7 isolates of A. flavus had 31% similarity. In this species, 6 isolates were in one group with two branches and only one isolate were resided in one branch similar to A. niger cluster. In addition, A. niger and A. flavus were 15 % similar.
Conclusion: Our results showed that the examined Aspergillus
species from air of different extents of distribution in Tehran had a significant genetic variation. Consequently, the genetic variation frequently happens among isolates of different areas. RAPD was concluded to be a useful technique for phylogenetic evaluation, which can be used as a complementary technique for differentiation of Aspergillus species when a little variation in morphological characters exists.
Keywords: Aspergillus, Outdoor air
P-25
Outdoor and indoor profile and seasonal variation of airborne Aspergilli spores in Shiraz (southern Iran) Hossein Khodadadi1, Ladan Karimi2, Mohammad Reza Samaei3, Fariba Abbasi3
1 Department of Medical Mycology and Parasitology, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran
2 Dr. Beheshti Hospital, Social Security Organization, Shiraz, Iran 3 Department of Environmental Health Engineering, School of Health, Shiraz University of Medical Sciences, Shiraz, Iran
Email: [email protected]
Introduction: Aspergillus species are important causes of respiratory disorders. However, data is limited regarding the diversity, load, seasonal distribution and environmental sources of airborne Aspergillus spores. Although recent studies suggest that Aspergillus flavus is a dominant clinical isolate in Iran, A. fumigatus is responsible for the majority of pulmonary aspergillosis cases in other regions of the world. This study aimed to describe the indoor and outdoor air levels and profiles of
Aspergillus spores and determine their associations in Shiraz, Iran.
Materials and Methods: Via According to the CDC bioaerosol sampling method (NIOSH 0800), 128 air samples were collected with a portable Andersen Air Impactor from different points of a compost facility, two hospitals and urban parts of Shiraz. Every point was sampled three times in summer and winter 2015. samples were cultured on Czapeck and Sabouraud chloramphenicol agar media. Aspergillus species were identified conventionally and viable Aspergillus CFU/m3 were counted. Differences between the selected environments were investigated statistically.
Results: In total, 125 Aspergillus species were isolated in this study, the foremost of which were A. flavus (38%), A. niger
(34%), A. fumigatus (26%), and Aspergillus spp. (2%), respectively. A. flavus was determined as the predominant species
in all regions during summer, while A. niger was the predominant species in the compost facility during winter. Level of A. fumigatus spores was mostly lower than the recommended quantity, while the concentrations of A. flavus and A. niger were extremely high in most outdoor sites. Seasonal variations of outdoor Aspergillus concentrations were affected by temperature, wind speed and humidity. Moreover, levels of Aspergillus spores were lower than the recommended amount in all indoor regions. Conclusion: Profiles and seasonal distribution patterns of
Aspergillus spores depend on the changes in atmospheric parameters (e.g., temperature, wind speed and humidity). In this study, profile patterns of airborne fungal spores were different due to the hot and dry climate of the southern region of Iran compared to areas with a moderate climate. Domination of environmental A. flavus spores in the majority of the studied regions could explain the higher isolation of A. flavus from Iranian patients compared to A. fumigatus. High concentration of airborne fungal spores in landfill sites is alarming. Therefore, fungal infections originated from outdoor environments must be prevented in workers and high-risk populations.
Keywords: Aspergillus, Spore levels, Outdoor, Seasonal variations
P-26
Coexistence of aspergilloma and pulmonary hydatid cyst Sadegh Khodavaisy1, Shirin Sadat Hashemi Fesharaki2,3, Hamid Badali3, Mohammadali Boroumand2, Maryam Sotoudeh Anvari2, Mohsen Gramishoar1,Zahra Abtahian4, Setareh Agha Kuchak Afshari1, Elahe Sasani1
1 Department of Medical Parasitology and Mycology, Tehran University of Medical Sciences, Tehran, Iran
2 Department of Pathology and Laboratory Medicine, Tehran Heart
Center, Tehran University of Medical Science, Tehran, Iran
3 Department of Medical Mycology and Parasitology, Invasive Fungi Research Center (IFRC), School of Medicine, Mazandaran University of Medical Sciences, Sari, Iran
4 Shahid beheshti university of medical sciences, Tehran, Iran Email: [email protected]
Introduction: Hydatid cyst is a zoonotic disease caused by
Echinococcus granulosus and Echinococcus multiloculari. The liver and lungs are the most common sites of infection although other organs are involved, as well. Pulmonary echinococcal hydatid cysts have been reported coexistent with aspergilloma. Case Presentation: In the current study, we report the case of successful treatment of the most disseminated coinfection of the hydatid cyst and aspergilloma due to Echinococcus granolosus
and Aspergillus flavus (as identifiedbased on molecular tools), respectively, in a 34-year-old female. In vitro antifungal susceptibility tests revealed that the minimum inhibitory concentrations for the antifungals used in this case in increasing order were posaconazole (0.031 µg/ml), itraconazole (0.125 µg/ml), voriconazole (0.25 µg/ml), and amphotericin B (1 µg/ml). The minimum effective concentration for caspofungin was 0.008 µg/ml.
Conclusion: This unique coexistence of active pulmonary echinococcosis and aspergillosis is reported because of its rarity and clinical importance for its management.
Keywords: Aspergilloma, Hydatid cyst, Aspergillusflavus
P-27
In vitro antifungal activities of amphotericin B, caspofungin, and fluconazole against Aspergillus terreus
Neda Kiasat1, Mahdis Zargaran1, Simin Taghipour1, Ali Zarei Mahmoudabadi1,2
1 Department of Medical Mycology, School of Medicine, Ahvaz
Jundishapur University of Medical Sciences, Ahvaz, Iran
2 Health Research Institute, Infectious and Tropical Diseases Research Center, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran
Email: [email protected]
among immunocompromised and high-risk patients. The prevalence of Aspergillus terreus, a cause of opportunistic infections (from superficial to serious invasive infections),is on a growing trend. Although invasive aspergillosis is often treated empirically with amphotericin B, most A. terreus isolates are drug resistant both in vivo and in vitro. The current study aimed to evaluate antifungals susceptibility profile of different environmental strains of Aspergillus terreus against amphotericin B, caspofungin, and fluconazole.
Materials and Methods:Forty A. terreus strains were isolated from environmental sources (air and soil) and identified, using macroscopic and microscopic features. Three antifungal drugs including amphotericin B, caspofungin, and fluconazole were applied for susceptibility test according to CLSI Broth Microdilution Method (M38-A2).
Results: The results of the study demonstrated that all the tested isolates had caspofungin MEC90 (4 μg/ml) higher than the epidemiological cutoff value, whereas only 20% (8) of the isolates exhibited amphotericin B MICs of ≤ 4 μg/ml. On the other hand, fluconazole showed a MIC90 of ≥ 128 μg/ml with a range of ≤ 1 to ≥ 128 μg/ml.
Conclusion: This study demonstrated that caspofungin had a better in vitro activity against all tested isolates of Aspergillus terreus, with MICs lower than two other antifu
